Subchronic memantine induced concurrent functional disconnectivity and altered ultra-structural tissue integrity in the rodent brain: revealed by multimodal MRI.
An effective NMDA antagonist imaging model may find key utility in advancing schizophrenia drug discovery research. We investigated effects of subchronic treatment with the NMDA antagonist memantine by using behavioural observation and multimodal MRI. Pharmacological MRI (phMRI) was used to map the neuroanatomical binding sites of memantine after acute and subchronic treatment. Resting state fMRI (rs-fMRI) and diffusion MRI were used to study the changes in functional connectivity (FC) and ultra-structural tissue integrity before and after subchronic memantine treatment. Further corroborating behavioural evidences were documented. Dose-dependent phMRI activation was observed in the prelimbic cortex following acute doses of memantine. Subchronic treatment revealed significant effects in the hippocampus, cingulate, prelimbic and retrosplenial cortices. Decreases in FC amongst the hippocampal and frontal cortical structures (prelimbic, cingulate) were apparent through rs-fMRI investigation, indicating a loss of connectivity. Diffusion kurtosis MRI showed decreases in fractional anisotropy and mean diffusivity changes, suggesting ultra-structural changes in the hippocampus and cingulate cortex. Limited behavioural assessment suggested that memantine induced behavioural effects comparable to other NMDA antagonists as measured by locomotor hyperactivity and that the effects could be reversed by antipsychotic drugs. Our findings substantiate the hypothesis that repeated NMDA receptor blockade with nonspecific, noncompetitive NMDA antagonists may lead to functional and ultra-structural alterations, particularly in the hippocampus and cingulate cortex. These changes may underlie the behavioural effects. Furthermore, the present findings underscore the utility and the translational potential of multimodal MR imaging and acute/subchronic memantine model in the search for novel disease-modifying treatments for schizophrenia.


Interaction of Bartonella henselae with endothelial cells results in rapid bacterial rRNA synthesis and replication.
Bartonella henselae is a slow-growing microorganism and the causative pathogen of bacillary angiomatosis in man. Here, we analysed how interaction of B. henselae with endothelial cells might affect bacterial growth. For this purpose, bacterial rRNA production and ribosome content was determined by fluorescence in situ hybridization (FISH) using rRNA-targeted fluorescence-labelled oligonucleotide probes. B. henselae grown on agar plates showed no detectable rRNA content by means of FISH, whereas B. henselae co-cultured with endothelial cells showed a rapid increase of rRNA production within the first 18 h after inoculation. The increased rRNA synthesis was paralleled by a approximately 1000-fold intracellular bacterial replication, whereas bacteria grown on agar base showed only a approximately 10-fold replication within the first 48 h of culture. Pretreatment of host cells with paraformaldehyde prevented adhesion, invasion, intracellular replication and bacterial rRNA synthesis of B. henselae. In contrast, inhibition of host cell protein synthesis by cycloheximide did not affect bacterial adhesion and invasion, but prevented intracellular replication although bacterial rRNA content was increased. Inhibition of actin polymerization by cytochalasin D did not affect adhesion, invasion, increased rRNA content or intracellular replication of B. henselae. These results demonstrate that rRNA synthesis and replication of B. henselae is promoted by viable host cells with intact de novo protein synthesis.


Spontaneously Arising Streptococcus mutans Variants with Reduced Susceptibility to Chlorhexidine Display Genetic Defects and Diminished Fitness.
Chlorhexidine (CHX) has been used to control dental caries caused by acid-tolerant bacteria such as Streptococcus mutans since the 1970s. Repeat CHX exposure for other bacterial species results in the development of variants with reduced susceptibility that also become more resistant to other antimicrobials. It has not been tested if such variants arise when streptococci are exposed to CHX. Here, we passaged S. mutans in increasing concentrations of CHX and isolated spontaneously arising reduced susceptibility variants (RSVs) from separate lineages that have MICs that are up to 3-fold greater than the parental strain. The RSVs have increased growth rates at neutral pH and under acidic conditions in the presence of CHX but accumulate less biomass in biofilms. RSVs display higher MICs for daptomycin and clindamycin but increased sensitivity to dental-relevant antimicrobials triclosan and sodium fluoride. Plate-based assays for competition with health-associated oral streptococci revealed decreased bacteriocin production by the RSVs, increased sensitivity to hydrogen peroxide, and diminished competitive fitness in a human-derived ex vivo biofilm consortium. Whole-genome sequencing identified common single nucleotide polymorphisms (SNPs) within a diacylglycerol kinase homolog and a glycolipid synthesis enzyme, which could alter the accumulation of lipoteichoic acids and other envelope constituents, as well as a variety of mutations in other genes. Collectively, these findings confirm that S. mutans and likely other streptococci can develop tolerance to CHX but that increased tolerance comes at a fitness cost, such that CHX-induced variants that spontaneously arise in the human oral cavity may not persist.


Support of kidney function by long-chain fatty acids derived from renal tissue.
To determine whether the oxidation of long-chain fatty acids (LCFA) derived from renal tissue lipids can support renal function, we perfused isolated rat kidneys with a substrate-free Krebs-Ringer bicarbonate solution containing 6 g/100 ml substrate-free (defatted) albumin. We measured GFR, TNa+, and Qo2 at 7-min intervals from 15 to 99 min after cannulation of the renal artery. Two groups (A and B) of 12 perfusions each were done. During substrate-free perfusion mean %TNa+ was low (A = 45 +/- 2%, B = 62 +/- 5%). When 10(-4) M 2-tetradecylglycidic acid (2-TDGA), a specific and irreversible inhibitor of long-chain acylcarnitine transferase-I, was added to the substrate-free perfusate, significant decreases in %TNa+ (A to approximately 25%; B to approximately 35%) and in Qo2 (delta = -25%) occurred. During perfusion with either 5 mM lactate or 5 mM alpha-ketoglutarate (alpha-KG) %TNa+ increased to approximately 80%. When 2-TDGA was added in the presence of lactate or of alpha-KG no decrease in %TNa+ or Qo2 occurred. Thus, 2-TDGA does not inhibit net renal Na+ transport or O2 uptake in the presence of high concentrations of lactate or alpha-KG, substrates not requiring long-chain acylcarnitine transferase for their utilization. We conclude that oxidation of LCFA released from renal tissue lipids can support a significant portion of Na+ reabsorption.


An indigenous conceptualization of reactive depression in Trinidad.
While depressive reactions appear to be universal, until recently they have seldom been described outside Europe, probably because of their lack of exotic salience, but also because they are seldom identified locally as distinct entities. Where it occurs, such an identification reflects a local preference for the articulation of individual psychological notions, rather than universal moral imperatives. While the symptoms of reactive depression are recognized in rural Trinidad as a common experience, but not especially remarked, they are also identified by Afro-Caribbeans as a specific state, Tabanka, following the loss of a sexual partner to another. Tabanka is characterized by lassitude, anorexia, insomnia, feelings of worthlessness, anger, a loss of interest in work and other activities and, especially, by a preoccupation with the faithless one. It is said to be most common among married men and among the upwardly socially mobile, and to lead to heavy drinking and occasionally madness or suicide. The precipitation of Tabanka provokes hilarity rather than moral indignation, and its humour articulates an egalitarian and individualistic working-class attitude to marriage and social mobility. In contrast, Trinidadians of Asian or Venezuelan origin emphasize moral notions of honour and shame.


Blast crisis in a murine model of chronic myelogenous leukemia.
The P210bcr/abl protein is produced in cells from patients with Philadelphia chromosome-positive chronic myelogenous leukemia (CML). Retroviral transfer of the gene encoding P210bcr/abl into murine bone marrow induces a granulocytic leukemia that models the chronic phase of human CML. We have transferred the leukemic clone to syngeneic animals, albeit with surprising inefficiency, and have observed CML and clonally related acute leukemias of lymphoid or myeloid phenotype in some transplant recipients. These data show that murine CML can result from retroviral transfer of the bcr/abl gene into pluripotent hematopoietic stem cells, that infected clones repopulate poorly after adoptive transfer, and that these clones can give rise to acute leukemia, reflecting evolution to a phase resembling blast crisis in the human disease.


Information propagated by longitudinal pulses near a van der Waals phase transition.
Longitudinal pulses that propagate in a medium near a van der Waals phase transition have a sigmoidal dependence on the strength of the stimulus arising from the phase structure. This response resembles the all-or-nothing property of action potentials, which raises the question if an acoustic system near a phase transition can be suitable for material-based neuromorphic computation. Herein, we investigate how information about the stimulus is stored within these pulses. We find that (1) the pulse propagates in parallel both digital and analog information about the stimulus amplitude; (2) the pulse encodes the type of stimulus, for instance, mechanical or thermal; and (3) a collision between two pulses stores information about both stimuli and may be used as a fading memory. Our results unravel a rich encoding of information in a phenomenon that is both common in a plethora of materials and mimics neuronal signaling. In addition, we show that these pulses carry more information than is typically considered by models of neural computation. Therefore, this phenomenon is an excellent candidate for in materio computation.


Diagnostic criteria of traumatic central cord syndrome. Part 3: descriptive analyses of neurological and functional outcomes in a prospective cohort of traumatic motor incomplete tetraplegics.
Prospective multicenter cohort study. To compare the neurological recovery and functional outcomes between traumatic central cord syndrome (TCCS) patients and motor incomplete tetraplegic patients. European Multicenter Study of human spinal cord injury. In 248 traumatic motor incomplete tetraplegics, initial phase (0-15 days) American Spinal Injury Association (ASIA) impairment grading, upper and lower extremity motor scores (UEMS and LEMS), upper and lower sensory scores and chronic phase (6 or 12 months) neurological outcomes were analyzed. In addition, chronic phase self-care and indoor mobility Spinal Cord Independence Measure (SCIM) items were studied. Tetraplegics were subdivided into three groups: (1) non-TCCS group (UEMSLEMS), (2) intermediate-TCCS group (UEMS=(1-9 points)<LEMS) and (3) TCCS group (UEMS=(10 points)<LEMS). Student's t-tests and χ(2)-tests were applied. A total of 89 non-TCCS subjects (AIS D, n=28), 62 int-TCCS (AIS D, n=43) and 97 TCCS (AIS D, n=80) subjects were analysed. Although minimal significant differences in chronic phase LEMS and UEMS outcomes were identified between TCCS and non-TCCS patients after stratification by the AIS grade, our data showed no significant differences in functional upper and lower extremity outcomes at 6 or 12 months post-injury. The AIS grading system, and not the diagnosis TCCS, continues to be the best available prognostic parameter for neurological and functional outcomes in motor incomplete tetraplegics. The authors recommend that for future outcome studies in motor incomplete tetraplegia, patients should not be selected based on, or stratified by, the diagnosis TCCS.


Four Decades of β-Lactam Antibiotic Pharmacokinetics in Cystic Fibrosis.
The pharmacokinetics (PK) of β-lactam antibiotics in cystic fibrosis (CF) patients has been compared with that in healthy volunteers for over four decades; however, no quantitative models exist that explain the PK differences between CF patients and healthy volunteers in older and newer studies. Our aims were to critically evaluate these studies and explain the PK differences between CF patients and healthy volunteers. We reviewed all 16 studies that compared the PK of β-lactams between CF patients and healthy volunteers within the same study. Analysis of covariance (ANCOVA) models were developed. In four early studies that compared adolescent, lean CF patients with adult healthy volunteers, clearance (CL) in CF divided by that in healthy volunteers was 1.72 ± 0.90 (average ± standard deviation); in four additional studies comparing age-matched (primarily adult) CF patients with healthy volunteers, this ratio was 1.46 ± 0.16. The CL ratio was 1.15 ± 0.11 in all eight studies that compared CF patients and healthy volunteers who were matched in age, body size and body composition, or that employed allometric scaling by lean body mass (LBM). Volume of distribution was similar between subject groups after scaling by body size. For highly protein-bound β-lactams, the unbound fraction was up to 2.07-fold higher in older studies that compared presumably sicker CF patients with healthy volunteers. These protein-binding differences explained over half of the variance for the CL ratio (p < 0.0001, ANCOVA). Body size, body composition and lower protein binding in presumably sicker CF patients explained the PK alterations in this population. Dosing CF patients according to LBM seems suitable to achieve antibiotic target exposures.


[Dyslipidemia and cardiovascular risk in type 2 diabetes mellitus patients with associated diabetic nephropathy].
Diabetes patients with concomitant diabetic nephropathy are especially destined to cardiovascular complications due to the presence of microalbuminuria or proteinuria, that are potent inductors of dyslipidaemia. We have studied 98 type 2 diabetes mellitus patients, 61 male and 37 female, mean age 63 +/- 13 year old, all of them with overt proteinuria (above 500 mg/day), divided into 4 groups: G-I (n = 13): patients with t. cholesterol > 6.25 mmol/l treated with fibric-acid derivatives; G-II (n = 52): hypercholesterolemic patients treated with statins; G-III (n = 20): hypercholesterolemic patients with no lipid-lowering intervention; G-IV (n = 13): normocholesterolemic patients (control group). Lipidic profile, proteinuria and renal function have being compared after 1, 3 and 5 years. Base-line characteristics of the patients were similar when regarding age, onset of diabetes or nephropathy. Only proteinuria was higher in statins-treated group (p < 0.05). Fibric-acid derivatives were more effective on hypertriglyceridaemia while statins were more effective lowering LDL cholesterol. A gemfibrocyl-treated patient presented a rhabdomyolysis episode. Statins were safe and well tolerated. Nine patients (19%) in G-II, 2 patients (10%) in G-III and 1 patient (7%) in G-IV achieved end-stage renal failure. Five-year cardiovascular mortality and all-cause mortality rate were 23%/23% in G-I, 13%/19% in G-II, 20%/25% in G-III and 31%/31% in G-IV. The difference was statistically significant when comparing normocolesterolemic versus statin-treated patients (p < 0.05). Lipid-lowering therapy could probably delay but not avoid the progression of diabetic nephropathy. Since dyslipidaemia is closely related to the progression of cardiovascular disease and mortality, an aggressive lipid-lowering therapy is recommended, irrespectively of its potential effect on diabetic nephropathy.


An economic analysis of chromosome testing in couples with children who have structural chromosome abnormalities.
Structural chromosome abnormalities can cause significant negative reproductive outcomes as they typically result in morbidity and mortality of newborns. The prevalence of structural chromosomal abnormalities in live births is at least 0.05%, of which many of them have parental origins. It is uncommon to predict structural chromosome abnormalities at birth in the first child but it is possible to prevent repeated abnormalities through screening and diagnostic programmes. This study will provide an economic analysis of the prenatal detection of these abnormalities. A cost-benefit analysis using a decision analytic model was employed to compare the status quo (doing nothing) with two interventional strategies. The first strategy (Strategy I) is preconceptional screening plus amniocentesis, and the second strategy (Strategy II) is amniocentesis alone. The monetary values in Thai baht (THB) were adjusted to international dollars (I$) using purchasing power parity (PPP) (I$1 = THB 17.60 for the year 2013). The robustness of the results was tested by applying a probabilistic sensitivity analysis. Both diagnostic strategies can reduce approximately 10.7-11.1 births with abnormal chromosomes per 1,000 diagnosed couples. The benefit cost ratios were 1.62 for Strategy I and 1.24 for Strategy II. Net present values per 1,000 diagnoses in couples were I$464,000 for Strategy I and I$267,000 for Strategy II. The probabilistic sensitivity analysis suggested that the cost-benefit analysis was sufficiently robust, confirming that both strategies provided higher benefits than costs. Since the benefits of both diagnostic strategies exceeded their costs, both strategies are economical-with Strategy I being more economically attractive. Strategy I is superior to Strategy II because it decreases the risk of normal children potentially dying from the amniocentesis process.


Inhibiting the initiation of Clostridium difficile spore germination using analogs of chenodeoxycholic acid, a bile acid.
To cause disease, Clostridium difficile spores must germinate in the host gastrointestinal tract. Germination is initiated upon exposure to glycine and certain bile acids, e.g., taurocholate. Chenodeoxycholate, another bile acid, inhibits taurocholate-mediated germination. By applying Michaelis-Menten kinetic analysis to C. difficile spore germination, we found that chenodeoxycholate is a competitive inhibitor of taurocholate-mediated germination and appears to interact with the spores with greater apparent affinity than does taurocholate. We also report that several analogs of chenodeoxycholate are even more effective inhibitors. Some of these compounds resist 7α-dehydroxylation by Clostridium scindens, a core member of the normal human colonic microbiota, suggesting that they are more stable than chenodeoxycholate in the colonic environment.


Continuous Directed Evolution of a Feedback-Resistant Arabidopsis Arogenate Dehydratase in Plantized Escherichia coli.
Continuous directed evolution (CDE) is a powerful tool for enzyme engineering due to the depth and scale of evolutionary search that it enables. If suitably controlled and calibrated, CDE could be widely applied in plant breeding and biotechnology to improve plant enzymes ex planta. We tested this concept by evolving Arabidopsis arogenate dehydratase (AtADT2) for resistance to feedback inhibition. We used an Escherichia coli platform with a phenylalanine biosynthesis pathway reconfigured ("plantized") to mimic the plant pathway, a T7RNA polymerase-base deaminase hypermutation system (eMutaT7), and 4-fluorophenylalanine as selective agent. Selection schemes were prevalidated using a known feedback-resistant AtADT2 variant. We obtained variants that had 4-fluorophenylalanine resistance at least matching the known variant and that carried mutations in the ACT domain responsible for feedback inhibition. We conclude that ex planta CDE of plant enzymes in a microbial platform is a viable way to tailor characteristics that involve interaction with small molecules.


Fused polypeptide with DEF induces apoptosis of lung adenocarcinoma cells.
To analyze the effects of a new unknown peptide DEF on the growth of tumor cells, a fused polypeptide TAT-DV1-DEF was designed and synthesized. The lung adenocarcinoma cell line GLC-82 treated with TAT- DV1-DEF was analyzed with a cell counting kit 8, and the location of polypeptides in cells was observed under laser confocal microscopy. The efficiency of polypeptide transfection and changes in nuclear morphology were analyzed by flow cytometry and fluorescence microscopy, respectively. Finally, the mechanism of tumor cell growth inhibition was evaluated by Western blotting. We found that TAT-DV1-DEF could significantly inhibit the growth of the lung adenocarcinoma cell line GLC-82, but not the normal human embryonic kidney cell line HEK-293. Polypeptides were found to be mostly localized in the cytoplasm and some mitochondria. The efficiency of polypeptide transfection in the two cell types was approximately 99%. Apoptotic nuclei were observed under fluorescence microscopy upon treatment with polypeptides and DAPI staining. Western blot analyses indicated that the polypeptide inhibition of tumor cell growth was apoptosis dependent. In the present study, we demonstrated that fused polypeptides could induce apoptosis of the lung adenocarcinoma cell line GLC-82, indicating that the new unknown peptide DEF has antitumor effects.


Encapsulation of Olive Pomace Extract in Rocket Seed Gum and Chia Seed Gum Nanoparticles: Characterization, Antioxidant Activity and Oxidative Stability.
The aim of this study was to determine the potential use of rocket seed and chia seed gum as wall materials, to encapsulate and to prevent degradation of olive pomace extract (OPE) in polymeric nanoparticles, and to characterize olive pomace extract-loaded rocket seed gum nanoparticles (RSGNPs) and chia seed gum nanoparticles (CSGNPs). The phenolic profile of olive pomace extract and physicochemical properties of olive pomace, rocket seed gum (RSG), and chia seed gum (CSG) were determined. The characterization of the nanoparticles was performed using particle size and zeta potential measurement, differential scanning calorimeter (DSC), Fourier-transform infrared spectroscopy (FTIR), scanning electron microscope (SEM), encapsulation efficiency (EE%), in vitro release, and antioxidant activity analyses. Nanoparticles were used to form oil in water Pickering emulsions and were evaluated by oxitest. The RSGNPs and CSGNPs showed spherical shape in irregular form, had an average size 318 ± 3.11 nm and 490 ± 8.67 nm, and zeta potential values of -22.6 ± 1.23 and -29.9 ± 2.57, 25 respectively. The encapsulation efficiency of the RSGNPs and CSGNPs were found to be 67.01 ± 4.29% and 82.86 ± 4.13%, respectively. The OPE-RSGNP and OPE-CSGNP presented peaks at the 1248 cm-1 and 1350 cm-1 which represented that C-O groups and deformation of OH, respectively, shifted compared to the OPE (1252.53 cm-1 and 1394.69 cm-1). The shift in wave numbers showed interactions of a phenolic compound of OPE within the RSG and CSG, respectively. In vitro release study showed that the encapsulation of OPE in RSGNPs and CSGNPs led to a delay of the OPE released in physiological pH. The total phenolic content and antioxidant capacity of RSGNPs and CSGNPs increased when the OPE-loaded RSGNPs and CSGNPs were formed. The encapsulation of OPE in RSGNPs and CSGNPs and the IP values of the oil in water Pickering emulsions containing OPE-RSGNPs and OPE-CSGNPs were higher than OPE, proving that OPE-loaded RSGNPs and CSGNPs significantly increased oxidative stability of Pickering emulsions. These results suggest that the RSG and CSG could have the potential to be utilized as wall materials for nanoencapsulation and prevent degradation of cold-pressed olive pomace phenolic extract.


Vascular Diameter as Clue for the Diagnosis of Clinically and/or Dermoscopically Equivocal Pigmented and Non-Pigmented Basal Cell Carcinomas and Nodular Melanomas.
Background and objectives: Dermoscopy is a useful tool for the early and non-invasive diagnosis of skin malignancies. Besides many progresses, heavily pigmented and amelanotic skin tumors remain still a challenge. We aimed to investigate by dermoscopy if distinctive morphologic characteristics of vessels may help the diagnosis of equivocal nodular lesions. Materials and Methods: A collage of 16 challenging clinical and dermoscopic images of 8 amelanotic and 8 heavily pigmented nodular melanomas and basal cell carcinomas was sent via e-mail to 8 expert dermoscopists. Results: Dermoscopy improved diagnostic accuracy in 40 cases. Vessels were considered the best clue in 71 cases. Focusing on the diameter of vessels improved diagnosis in 5 cases. Conclusions: vascular diameter in addition to morphology and arrangement may be a useful dermoscopic clue for the differential diagnosis of clinically equivocal nodular malignant tumors.


[Malignant tumor mortality in groups of female medical workers].
The paper discusses results of a descriptive epidemiologic study of cancer mortality in the following three groups of retired women: (1) physicians of all specialties, (2) physician's assistants, midwives and nurses, and (3) practical nurses. Cancer mortality proved significantly increased in all the groups studied: 1.5, 1.3 and 1.1 higher, respectively. Moreover, the physician group was characterized by significantly higher mortality from rectal cancer, breast cancer and hemoblastoses (3.1, 4.2 and 2.6 times higher, respectively). Mortality rates for lung and breast cancer were 1.6 and 1.2 times higher in the nurse group whereas that for cervical cancer was increased in the practical nurses.


Artificial compound eye applying hyperacuity.
Inspired by the natural phenomenon of hyperacuity, redundant sampling in combination with the knowledge about the impulse response of the imaging system is used to extract highly accurate information using a low resolving artificial apposition compound eye. Thus the implementation of a precise position detection for simple objects like point sources and edges is described.


Role of heme in the regulation of human and rat platelet soluble guanylate cyclases.
The chromatography of soluble human and rat platelet guanylate cyclases (105000 g supernatants) on DEAE-cellulose in 50 mM Tris HCl buffer, containing 0.22 M NaCl, has yielded virtually identical elution profiles, each with two protein peaks (I and II). Only peak II was found to have guanylate cyclase activity. Experiments with human platelets showed that inactive protein peak I inhibited the activity of guanylate cyclase preparation (peak II) and restored the already lost ability of the enzyme to be activated by sodium nitroprusside. In experiments with rat platelets, inactive fraction I had no effect on guanylate cyclase activity (peak II), and the enzyme was not activated by sodium nitroprusside either before or after DEAE-cellulose. 105000g supernatant of human platelets had an absorbance maximum at 415 nm (Soret band), which disappeared from the spectrum of the active fraction (II) but was found in the spectrum of the inactive (inhibitory) fraction I. Experiments with rat platelets demonstrated the absence of Soret band in the corresponding spectra. It was concluded that, contrary to the generally accepted notion, heme is not a prosthetic group of the soluble rat platelet guanylate cyclase.


The coordinate replication of the human beta-globin gene domain reflects its transcriptional activity and nuclease hypersensitivity.
The temporal order of replication of DNA sequences in the chromosomal domain containing the human beta-globin gene cluster and its flanking sequences (140 kilobases) was measured and compared in two different human cell lines. In human erythroleukemia (K562) cells, in which embryonic and fetal globin genes are transcribed, all of the sequences we examined from the beta-globin domain replicated early during S phase, while in HeLa cells, in which globin genes are transcriptionally silent, these sequences replicated late during S. Potential sites of initiation of DNA replication within this domain were identified. The beta-globin gene domain was also found to differ with respect to the nuclease sensitivity of the chromatin in these two cell lines. In K562 cells, hypersensitive sites for endogenous nucleases and DNase I were present in the chromatin near the earliest-replicating segments in the beta-globin domain.


Monitoring psychrotrophic lactic acid bacteria contamination in a ready-to-eat vegetable salad production environment.
A study monitoring lactic acid bacteria contamination was conducted in a company producing fresh, minimally processed, packaged and ready-to-eat (RTE) vegetable salads (stored at 4°C) in order to investigate the reason for high psychrotrophic LAB levels in the products at the end of shelf-life. Initially, high microbial counts exceeding the established psychrotrophic thresholds (>10(7)-10(8)CFU/g) and spoilage manifestations before the end of the shelf-life (7days) occurred in products containing an assortment of sliced and diced vegetables, but within a one year period these spoilage defects became prevalent in the entire processing plant. Environmental sampling and microbiological analyses of the raw materials and final products throughout the manufacturing process highlighted the presence of high numbers of Leuconostoc spp. in halved and unseeded, fresh sweet bell peppers provided by the supplier. A combination of two DNA fingerprinting techniques facilitated the assessment of the species diversity of LAB present in the processing environment along with the critical point of their introduction in the production facility. Probably through air mediation and surface adhesion, mainly members of the strictly psychrotrophic species Leuconostoc gelidum subsp. gasicomitatum and L. gelidum subsp. gelidum were responsible for the cross-contamination of every vegetable handled within the plant.


Serogrouping of Bacteroides nodosus isolates from 62 sources in the United States.
Bacteroides nodosus isolates from 62 sources in the United States were obtained from sheep with infectious foot diseases. Serotypic analysis of these isolates revealed 21 serotypes (designated I-XXI). These serotypes were compared with British and Australian/New Zealand B nodosus strains by use of reciprocal tube agglutination tests. These tests, as well as the cross-matching tube agglutination tests of the US serotypes, resulted in arranging the US serotypes into 11 serogroups, and comparing these serogroups with their Australian/New Zealand serogroup and British serotype counterparts. Three US serogroups and 1 additional British serotype had little or no relationship to any of the Australian/New Zealand serogroups A-H (the vaccine strains). One or more of these unrelated serogroups were found in 29% of the sources studied. The most frequently found US serotype was serotype XV at 29%. The most frequently found US serogroups were the serogroups analogous to serogroup B (43.5%) and serogroup H (37%); the other serogroups were found in 22.6% or less of the sources studied. Evaluation of 3 sources revealed that multiple serotypes in a single flock are common, multiple serotypes from a single lesion are possible, B nodosus isolates obtained from goats (unlike those from cattle) appear identical to the isolates obtained from sheep, and disease can appear in vaccinated animals, even in a flock that appears to be harboring only a single serogroup-B serotype (the serogroup for which there are 3 strains in the current vaccine).


The effect of spinal analgesia on visceral nociceptive neurons in caudal medulla of the rat.
A population of neurons resident in the caudal ventrolateral medulla are excited by noxious cutaneous and visceral stimuli from large portions of the body. These neurons act as monitors of ascending nociceptive information, and we hypothesized that they would be inhibited by spinally administered analgesics in a clinically relevant fashion. Rats were anesthetized with oxygen/ halothane. The caudal medulla was surgically exposed, and a catheter placed into the intrathecal space overlying the lower thoracic spinal cord via the surgical site. Single medullary neurons were characterized for responses to cutaneous and visceral (colorectal distension) stimuli. The effects of i.v. and intrathecally administered morphine and lidocaine were determined. The intrathecal infusion of morphine for 6 days before testing was also used as a pretreatment. Colorectal distension-evoked responses of medullary nociceptive neurons were inhibited in a dose-dependent, naloxonereversible fashion by intrathecal and i.v. morphine (50% effective dose values: 3.5 and 440 microg/kg, respectively). Intrathecal lidocaine abolished responses to colorectal distension and produced a spinal level at doses producing minimal effects when administered systemically. Prior treatment with an infusion of morphine produced tolerance to the effects of subsequent intrathecal morphine administration. These findings support the use of this preparation as a neurophysiologic model of spinal analgesia. Neurons in the brainstem, isolated electrophysiologically, were used as whole body monitors of pain-related activity in the rat. As a neurophysiologic model of nociception, this preparation may prove useful for the study of regionally administered analgesics and local anesthetics.


Transitions and turning points revisited: A replication to explore child maltreatment and youth offending links within and across Australian cohorts.
In this study we examined the consistency of life-course child maltreatment trajectories and youth offending links across birth cohorts. In so doing we demonstrated the value of replication studies for maltreatment research. We applied the methodology of Stewart et al. (2008) and linked population-based (1990 birth cohort) child protection and youth justice administrative data from Queensland, Australia. We performed a group based trajectory analysis to identify distinct maltreatment trajectory groups distinguishable by maltreatment timing and frequency across the life-course. We explored group-based youth offending outcomes with consideration of variations in maltreatment chronicity, timing, and frequency, multi-type maltreatment, gender and race (Indigenous Australian versus non-Indigenous Australian youths). To determine the consistency of maltreatment trajectories and offending links across cohorts (1983/84 versus 1990) we compared our results with those of Stewart et al. (2008). Consistent with Stewart et al. (2008): (1) We identified six distinct maltreatment trajectory groups; (2) Trajectory groups characterised by chronic maltreatment and/or adolescent maltreatment had the largest proportion of young offenders; and (3) Maltreatment frequency commonly peaked at transition points. Extending beyond Stewart et al. (2008) we noted considerable overlap between maltreatment dimensions and a potential impact of race and multi-type maltreatment on maltreatment and offending links. We endorse replication studies as a valuable tool to advance child maltreatment policy and practice and recommend further research on interactions between maltreatment dimensions, gender, race, and youth offending.


Vaginectomy: a minimally invasive treatment for cervical cancer vaginal recurrence.
Radiotherapy and/or pelvic exenteration represent the treatment of vaginal recurrence, but the prognosis remains unsatisfactory and with long-term complications. We investigated the possible role of vaginectomy for isolated vaginal relapse (IVR) in cervical cancer (CC). Patients with vaginal CC recurrence were evaluated for surgical treatment consisting in vaginectomy. Data were prospectively collected and analyzed to identify independent prognostic factors. Twenty-nine patients with IVR from CC were enrolled. Early and late complications were observed in 7 (24%) and 6 (21%) patients, respectively. After a median follow-up of 57.5 months (range, 8-100 months), 16 patients (55%) were disease-free. The 5-year overall survival and progression-free survival rates were 70.5% and 59.4%, respectively. In carefully selected patients, vaginectomy may be considered a therapeutic option for IVR. Older patients with long disease-free interval and small recurrences benefit the most from this bladder-sparing surgical technique.


Molecular isoforms of the sodium pump in brain.
The Na+,K+-ATPase is a basic membrane element in all animal cells that maintain transmembrane gradients of Na+ and K+ ions. When integrated into artificial lipid vesicles the isolated Na+,K+-ATPase is capable of actively transporting Na+ and K+, thus establishing its identity with the well-known Na+ pump. Three isoforms of the catalytic subunit of Na+,K+-ATPase have recently been identified in the rat brain. Elucidating the regulatory processes which govern their biosynthesis in the brain is central to understanding the physiological alterations which accompany diabetes, chronic cardiotonic steroid intoxication and degenerative diseases of the retina. In this study, synthesis of the alpha-1 and alpha-2 isoforms was examined in the retina of rats following intraocular injection of citrate buffer. Incorporation of [35S]methionine into both isoforms was increased 60-100% by 6 hr, demonstrating that neuronal Na+ pump is subject to regulation, although the question of independent regulatory controls remains unanswered.


Gender differences in Ca(2+) entry mechanisms of vasoconstriction in Wistar-Kyoto and spontaneously hypertensive rats.
We investigated whether putative vascular protection against hypertension in females reflects differences in the Ca(2+) mobilization mechanisms of vasoconstriction depending on the gender and the status of the gonads. Active stress and (45)Ca(2+) influx were measured in aortic strips isolated from intact and gonadectomized male and female Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR). In aortic strips of intact male WKY incubated in normal Krebs' solution (2.5 mmol/L Ca(2+)), both phenylephrine (10(-5) mol/L) and membrane depolarization by 96 mmol/L KCl caused significant increases in active stress and (45)Ca(2+) influx. In intact female WKY, the phenylephrine- and KCl-induced stress and (45)Ca(2+) influx were significantly reduced. In Ca(2+)-free (2 mmol/L EGTA) Krebs' solution, stimulation of aortic strips with phenylephrine or caffeine (25 mmol/L) to induce Ca(2+) release from intracellular stores caused a transient increase in stress that was not significantly different between males and females. In SHR, the phenylephrine- and KCl-induced stress and (45)Ca(2+) influx were significantly greater than those in WKY in all groups of rats. The reduction in stress and Ca(2+) entry in intact females compared with intact males was greater in SHR than in WKY. The contractile responses and Ca(2+) entry in castrated male and ovariectomized female WKY or SHR were not significantly different from the respective responses in intact males. The contractile responses and Ca(2+) entry in ovariectomized female WKY or SHR with 17beta-estradiol implant were not significantly different from the respective responses in intact females. Thus, the phenylephrine- and depolarization-induced vascular reactivity and Ca(2+) entry in vascular smooth muscle are dependent on gender and on the presence or absence of functional female gonads. Ca(2+) release from intracellular stores is not affected by gender or gonadectomy. The gender-specific changes in vascular reactivity and Ca(2+) entry are augmented in hypertension.


[Monitoring of implementation of international programs of poliomyelitis eradication and measles and rubella elimination in the Republic of Belarus].
Monitoring of implementation of international programs of poliomyelitis eradication, and measles and rubella elimination in the Republic of Belarus based on results of molecular-epidemiologic studies of 2009 - 2010. 271 viral agents isolated from children with acute flaccid paralysis syndrome, other diseases, healthy children and from sewage water within the framework of poliomyelitis control implementation were identified by serological and molecular methods. Blood sera of 528 patients with fever and rash were examined for the presence of IgM to measles and rubella virus, 418 - for the presence of IgM to parvovirus B19 and parvovirus DNA. Blood sera of 33 pregnant women and 64 children with signs of intrauterine infection were studied for IgM and IgG antibodies to rubella virus. Measles virus was isolated, N-gene sequence and phylogenetic analysis carried out. The studies performed confirmed that indigenous wild polioviruses in the country do not circulate, imported wild or vaccine-related polioviruses were also not detected. Measles and rubella morbidity in the Republic of Belarus was less than 1 in 1 000 000. 2 cases of rubella (2009) and 1 case of measles (2010) was detected during adequate control level: the rate of detection of patients with fever and rash, in whom measles and rubella diagnosis was excluded by the results of laboratory examination, was more than 2 in 100 000 of the population. The etiologic agent in more than 20% of diseases with fever and rash was parvovirus B19. A single case of measles was caused by genotype D8 virus imported from India. The data obtained give evidence to conformance of the poliomyelitis, measles, rubella, innate rubella syndrome control implemented in the Republic of Belarus to WHO recommendations; maintenance of status of country as free from poliomyelitis and achievement of main criteria of elimination of both measles and rubella by 2010.


Expression of creatine kinase isoenzyme genes during postnatal development of rat brain cerebellum: evidence for transcriptional regulation.
Transcription and accumulation of brain-type creatine kinase (CKB) mRNA and its protein was examined during postnatal development of rat brain cerebellum, the brain region containing highest CKB mRNA in the adult. CKB protein was extremely low at day 1, increased about 10-fold until week 4 and remained constant until week 10. This time course was paralleled by cerebellar CKB mRNA, which was also extremely low at day 1 and increased 5-fold during the first 3 weeks and then remained constant. High levels of CKB protein were also detected in cultured primary cerebellar granular neurons. Nuclear run-on assays directly showed that CKB mRNA accumulation during postnatal cerebellar development was due to increased transcription. When compared with cerebrum and whole brain, cerebellar CKB mRNA accumulation during postnatal development was temporally delayed. Analysis of myocyte enhancer factor (MEF)-2 and Sp1, factors known to initiate or sustain CKB transcription in tissues other than brain, revealed that MEF-2 in cerebellum was low at week 1 but increased 3.5-fold by week 7, while Sp1 remained unchanged. The increase in CKB protein during cerebellar postnatal development was coincident with that of the ubiquitous mitochondrial CK protein and mRNA, indicating that a functional phosphocreatine energy shuttle probably exists for efficient ATP regeneration in the cerebellum. This should be beneficial for the many energy-demanding requirements during cerebellar development, as indicated by the observed temporal co-expression of CKB with myelin basic protein, which is involved in axon myelination by oligodendrocytes.


Application of real-time quantitative PCR assays for detecting marine Brucella spp. in fish.
Brucella ceti and Brucella pinnipedialis have been documented as occurring in marine mammals, and B. ceti has been identified in 3 naturally acquired human cases. Seroconversion and infection patterns in Pacific Northwest harbor seals ( Phoca vitulina richardii) and North Atlantic hooded seals ( Cystophora cristata) indicate post-weaning exposure through prey consumption or lungworm infection, suggesting fish and possibly invertebrates play an epizootiologic role in marine Brucella transmission and possible foodborne risk to humans. We determined if real-time quantitative PCR (qPCR) assays can detect marine Brucella DNA in fish DNA. Insertion sequence (IS) 711 gene and sequence type (ST)27 primer-probe sets were used to detect Brucella associated with marine mammals and human zoonotic infections, respectively. First, DNA extracts from paired-species fish (containing 2 species) samples were tested and determined to be Brucella DNA negative using both IS 711 and ST27 primer-probe sets. A representative paired-species fish DNA sample was spiked with decreasing concentrations of B. pinnipedialis DNA to verify Brucella detection by the IS 711 primer-probe within fish DNA. A standard curve, developed using isolated DNA from B. pinnipedialis, determined the limit of detection. Finally, the IS 711 primer-probe was used to test Atlantic cod ( Gadus morhua) DNA extracts experimentally infected with the B. pinnipedialis hooded seal strain. In culture-positive cod tissue, the IS 711 limit of detection was ~1 genome copy of Brucella. Agreement between culture and PCR results for the 9 positive and 9 negative cod tissues was 100%. Although a larger sample set is required for validation, our study shows that qPCR can detect marine Brucella in fish.


Current understanding of pathogenetic mechanisms in neuroendocrine neoplasms.
Despite the fact that important advances in research on neuroendocrine neoplasms (NENs) have been made, consistent data about their pathogenetic mechanism are still lacking. Furthermore, different primary sites may recognize different pathogenetic mechanisms. This review analyzes the possible biological and molecular mechanisms that may lead to NEN onset and progression in different organs. Through extensive research of the literature, risk factors including hypercholesterolemia, inflammatory bowel disease, chronic atrophic gastritis are evaluated as potential pathogenetic mechanisms. Consistent evidence is available regarding sporadic gastric NENs and MEN1 related duodenopancreatic NENs precursor lesions, and genetic-epigenetic mutations may play a pivotal role in tumor development and bone metastases onset. In lung neuroendocrine tumors (NETs), diffuse proliferation of neuroendocrine cells on the bronchial wall (DIPNECH) has been proposed as a premalignant lesion, while in lung neuroendocrine carcinoma nicotine and smoke could be responsible for carcinogenic processes. Also, rare primary NENs such as thymic (T-NENs) and Merkel cell carcinoma (MCC) have been analyzed, finding different possible pathogenetic mechanisms. New technologies in genomics and epigenomics are bringing new light to the pathogenetic landscape of NENs, but further studies are needed to improve both prevention and treatment in these heterogeneous neoplasms.


Chronic hypersensitivity for inflammatory nociceptor sensitization mediated by the epsilon isozyme of protein kinase C.
We have identified a mechanism, mediated by the epsilon isozyme of protein kinase C (PKCepsilon) in peripheral neurons, which may have a role in chronic inflammatory pain. Acute inflammation, produced by carrageenan injection in the rat hindpaw, produced mechanical hyperalgesia that resolved by 72 hr. However, for up to 3 weeks after carrageenan, injection of the inflammatory mediators prostaglandin E(2) or 5-hydroxytryptamine or of an adenosine A(2) agonist into the same site induced a markedly prolonged hyperalgesia (>24 hr compared with 5 hr or less in control rats not pretreated with carrageenan). A nonselective inhibitor of several PKC isozymes and a selective PKCepsilon inhibitor antagonized this prolonged hyperalgesic response equally. Acute carrageenan hyperalgesia could be inhibited by PKA or PKG antagonists. However, these antagonists did not inhibit development of the hypersensitivity to inflammatory mediators. Our findings indicate that different second messenger pathways underlie acute and prolonged inflammatory pain.


Cap analogs containing 6-thioguanosine--reagents for the synthesis of mRNAs selectively photo-crosslinkable with cap-binding biomolecules.
Numerous biomolecules recognize the 7-methylguanosine cap structure present at the 5' ends of eukaryotic mRNAs. Photo-crosslinking is a valuable technique to study these interactions. We report three anti-reverse cap analogs containing a photo-activable nucleoside, 6-thioguanosine ((6S)G), that enable the synthesis of capped RNAs with (6S)G positioned exclusively as the first transcribed nucleotide. The effect of the 6-thioguanosine moiety on binding to the translation factor eIF4E and the efficiency of mRNA translation was determined. The utility of mRNAs with a (6S)G-modified cap in crosslinking experiments is shown by mapping the histone H4 cap-binding pocket.


The role of indocyanine green videoangiography with FLOW 800 analysis for the surgical management of central nervous system tumors: an update.
OBJECTIVE Indocyanine green videoangiography (ICG-VA) is an intraoperative technique used to highlight vessels in neurovascular surgery. Its application in the study of the vascular pathophysiology in CNS tumors and its role in their surgical management are still rather limited. A recent innovation of ICG-VA (i.e., the FLOW 800 algorithm integrated in the surgical microscope) allows a semiquantitative evaluation of cerebral blood flow. The aim of this study was to evaluate for the first time the systematic application of ICG-VA and FLOW 800 analysis during surgical removal of CNS tumors. METHODS Between May 2011 and December 2017, all cases in which ICG-VA and FLOW 800 analysis were used at least one time before, during, or after the tumor resection, and in which surgical videos were available, were retrospectively reviewed. Results of the histological analysis were analyzed together with the intraoperative ICG-VA with FLOW 800 in order to investigate the tumor-related videoangiographic features. RESULTS Seventy-one patients who underwent surgery for cerebral and spinal tumors were intraoperatively analyzed using ICG-VA with FLOW 800, either before or after tumor resection, for a total of 93 videoangiographic studies. The histological diagnosis was meningioma in 25 cases, glioma in 14, metastasis in 7, pineal region tumor in 5, hemangioblastoma in 4, chordoma in 3, and other histological types in 13 cases. The authors identified 4 possible applications of ICG-VA and FLOW 800 in CNS tumor surgery: extradural surveys allowed exploration of sinus patency and the course of veins before dural opening; preresection surveys helped in identifying pathological vascularization (arteriovenous fistulas and neo-angiogenesis) and regional venous outflow, and in performing temporary venous clipping tests, when necessary; postresection surveys were conducted to evaluate arterial and venous patency and parenchymal perfusion after tumor removal; and a premyelotomy survey was conducted in intramedullary tumors to highlight the posterior median sulcus. CONCLUSIONS The authors found ICG-VA with FLOW 800 to be a useful method to monitor blood flow in the exposed vessels and parenchyma during microsurgical removal of CNS tumors in selected cases. In particular, a preresection survey provides useful information about pathophysiological changes of brain vasculature related to the tumor and aids in the individuation of helpful landmarks for the surgical approach, and the postresection survey helps to prevent potential complications associated with the resection (such as local hypoperfusion or venous infarction).


Ultra-Short Race-Pace Training (USRPT) In Swimming: Current Perspectives.
The last decade has seen a dramatic rise in sports science research due to the ever-increasing professionalization of sport. As a result, many alternative training methodologies that challenge traditional training philosophies have emerged. In the sport of swimming, ultra-short race-pace training (USRPT) was recently proposed. The aim of this article was to provide current perspectives on USRPT in competitive swimming. A systematic review was conducted to determine the effects of USRPT on performance in competitive swimmers. Of the 1347 studies retrieved, 1332 were excluded. The full-texts of 15 studies were assessed for eligibility. However, all 15 studies were excluded as the intervention did not consist of USRPT. Consequently, there are concerns surrounding USRPT as it is not currently based on peer-reviewed published literature. In addition, the recommendations within USRPT to avoid resistance training, cross-training activities, training intensities less than race-pace velocity and part practice swimming drills are highly controversial and lack scientific evidence. There is evidence to suggest that USRPT is a derivative of high-intensity training (HIT) and there is peer-reviewed published literature available to support the effects of HIT on performance in competitive swimmers. Swimming coaches and sports scientists are advised to consider the applications of USRPT with caution. The authors suggest that USRPT is a training method, which may be incorporated within a holistic periodized training program that includes a variety of training methods and stimuli. Future research should involve a randomized controlled intervention of USRPT in competitive swimmers.


[Medical problems of handedness].
Handedness is one example of many forms of behavioral lateralization seen in humans. In the present review the practical relevance and the problems of left-handedness are discussed. Left-handedness has existed in a small subset of the human population, approximately 8%, since the origin of man. The incidence of left-handedness as usually defined is consistent among human populations. An understanding of handedness may lead to valuables clues as to how the brain becomes organised in the way it does. Several authors suggested a pathological handedness explaining the origin of laterality in man. There is a general agreement that sinistrality can occur as a result of pre- or perinatal damage to the left hemisphere. A disproportionate number of sinistrals is found in clinical populations. The review indicates that there is no sure evidence to suggest that left-handedness is a pathological sign.


[Is it necessary to remove polycystic kidneys?].
The Multicystic Dysplastic Kidney is the most frequent renal congenital anomaly and the second most common cause of an abdominal mass in the newborn. Traditionally, since this pathological entity was described as such by Spene in 1955, early resection has been deemed its best treatment. In the last decade, a conservative approach has been advocated by some authors, except for the patients presenting with concomitant abdominal pain or infection. In our hospital, relying on the support afforded by the echographic assessment of these patients, we have proposed an expectant behaviour in the management of this entity. So far, we have had the opportunity to follow the course of the disease in 15 patients with the diagnosis of Multicystic Dysplastic Kidney, with the result that in no one of them the size of the cystic mass increased, but instead the mass disappeared completely in one case, diminished greatly in three cases, had a moderate decrease in size in eight cases and in just three patients it has not varied in size. No patient has complained of abdominal pain or has become infected. In the face of such findings, we believe that early resection of the multicystic kidney is not warranted if no complications develop.


Prediction of intradialytic hypotension using photoplethysmography.
Intradialytic hypotension is the most common acute complication during conventional hemodialysis treatment. Prediction of such events is highly desirable in clinical routine for prevention. This paper presents a novel prediction method of acute symptomatic hypotension in which the photoplethysmographic signal is analyzed with respect to changes in amplitude, reflecting vasoconstriction, and cardiac output. The method is based on a statistical model in which the noise is assumed to have Laplacian amplitude distribution. The performance is evaluated on 11 hypotension-prone patients who underwent hemodialysis treatment, resulting in seven events with acute symptomatic hypotension and 17 without. The photoplethysmographic signal was continuously acquired during treatment as was information on blood pressure and oxygen saturation. Using leave-one-out cross validation, the proposed method predicted six out of seven hypotensive events, while producing 1 false prediction out of 17 possible. The performance was achieved when the prediction threshold was chosen to be in the range 57%-65% of the photoplethysmographic envelope at treatment onset.


Activation mechanism and function of the MAP kinase cascade.
MAP kinase (MAPK) and its activator, MAP kinase kinase (MAPKK), are commonly activated by a variety of extracellular stimuli in mammalian cells and in the process of Xenopus oocyte maturation. In order to investigate the function of the MAPK cascade in oocyte maturation, we produced an anti-Xenopus MAPKK which specifically reacts with MAPKK in vitro. When this antibody was microinjected into immature oocytes, MAPK activation induced by progesterone was prevented. Surprisingly, H1 kinase activation and germinal vesicle breakdown were also inhibited in the oocytes injected with this antibody. These results suggest that the MAPK cascade plays an important role in the maturation promoting factor (MPF) activation during the oocyte maturation process. When this antibody together with Mos was microinjected into Xenopus two-cell embryos, the Mos-induced metaphase arrest (CSF arrest) was prevented. Thus, the MAPK cascade may mediate CSF arrest. During Xenopus early embryogenesis, a low but significant level of MAPK remains active. Injection of mRNA encoding a constitutively active MAPKK resulted in mesoderm induction in animal cap explants. In addition, fibroblast growth-factor (FGF)-induced mesoderm induction was inhibited by expressing CL100 (a MAP kinase phosphatase) in animal cap explants. Thus the MAPK cascade may be involved in the mesoderm induction of Xenopus embryos. The activation pathways and roles of the MAPKK/MAPK cascade in various signaling processes will be discussed.


IKK beta is required for peripheral B cell survival and proliferation.
NF-kappaB activity in mammalian cells is regulated through the IkappaB kinase (IKK) complex, consisting of two catalytic subunits (IKKalpha and IKKbeta) and a regulatory subunit (IKKgamma). Targeted deletion of Ikkbeta results in early embryonic lethality, thus complicating the examination of IKKbeta function in adult tissues. Here we describe the role of IKKbeta in B lymphocytes made possible by generation of a mouse strain that expresses a conditional Ikkbeta allele. We find that the loss of IKKbeta results in a dramatic reduction in all peripheral B cell subsets due to associated defects in cell survival. IKKbeta-deficient B cells are also impaired in mitogenic responses to LPS, anti-CD40, and anti-IgM, indicating a general defect in the ability to activate the canonical NF-kappaB signaling pathway. These findings are consistent with a failure to mount effective Ab responses to T cell-dependent and independent Ags. Thus, IKKbeta provides a requisite role in B cell activation and maintenance and thus is a key determinant of humoral immunity.


Gene expression profiling revealed novel molecular targets of docetaxel and estramustine combination treatment in prostate cancer cells.
Both docetaxel and estramustine are antimicrotubule agents with antitumor activity in various cancers including prostate cancer. Clinical trials for docetaxel and estramustine combination treatment have suggested improved antitumor activity in hormone-refractory prostate cancer. However, the molecular mechanisms involved in the combination treatment with docetaxel and estramustine have not been fully elucidated. In order to establish such molecular mechanisms in both hormone insensitive (PC-3) and sensitive (LNCaP) prostate cancer cells, gene expression profiles of docetaxel- and estramustine-treated prostate cancer cells were obtained by using Affymetrix Human Genome U133A Array. Total RNA from PC-3 and LNCaP cells untreated and treated with 2 nmol/L docetaxel, 4 micromol/L estramustine, or 1 nmol/L docetaxel plus 2 micromol/L estramustine for 6, 36, and 72 hours was subjected to microarray analysis. Real-time PCR and Western blot analysis were conducted to confirm the microarray data. Clustering analysis based on biological function showed that docetaxel and estramustine combination treatment down-regulated some genes that are known to regulate cell proliferation, transcription, translation, and oncogenesis. In contrast, docetaxel and estramustine combination treatment up-regulated some genes related to induction of apoptosis, cell cycle arrest, and tumor suppression. Docetaxel and estramustine also showed differential effects on gene expression between mono- and combination treatment. Combination treatment with docetaxel and estramustine caused alternations of a large number of genes, many of which may contribute to the molecular mechanisms by which docetaxel and estramustine inhibit the growth of prostate cancer cells. These results provide novel molecular targets of docetaxel and estramustine combination treatment in prostate cancer cells. This information could be utilized for further mechanistic research and for devising optimized therapeutic strategies against prostate cancer.


Evaluation of a pump in enteral tube feeding in terms of patient and staff acceptance: a retrospective study.
A retrospective study of 39 patients fed via an enteral tube was carried out using data from patients records. An enteral feeding pump was available for all patients. Its use was evaluated in terms of patient and staff acceptance of the technique. The pump was shown to be particularly suited to patients following gastrointestinal surgery and trauma in whom tube feeding might be expected to produce gastrointestinal side-effects. Patients with lower nutritional requirements and without gastrointestinal trauma do not require such accurate flow control and for non-clinical reasons an alternative technique such as gravity drip feed or bolus feeding may be preferred. The conclusions which can be drawn from a study of this nature are limited but it does draw attention to the importance of considering, in addition to the expected clinical tolerance, the acceptance by patient and staff of the technique for the individual.


A fifth major genetic group among honeybees revealed in Syria.
Apiculture has been practiced in North Africa and the Middle-East from antiquity. Several thousand years of selective breeding have left a mosaic of Apis mellifera subspecies in the Middle-East, many uniquely adapted and survived to local environmental conditions. In this study we explore the genetic diversity of A. mellifera from Syria (n = 1258), Lebanon (n = 169) and Iraq (n = 35) based on 14 short tandem repeat (STR) loci in the context of reference populations from throughout the Old World (n = 732). Our data suggest that the Syrian honeybee Apis mellifera syriaca occurs in both Syrian and Lebanese territories, with no significant genetic variability between respective populations from Syria and Lebanon. All studied populations clustered within a new fifth independent nuclear cluster, congruent with an mtDNA Z haplotype identified in a previous study. Syrian honeybee populations are not associated with Oriental lineage O, except for sporadic introgression into some populations close to the Turkish and Iraqi borders. Southern Syrian and Lebanese populations demonstrated high levels of genetic diversity compared to the northern populations. This study revealed the effects of foreign queen importations on Syrian bee populations, especially for the region of Tartus, where extensive introgression of A. m. anatolica and/or A. m. caucasica alleles were identified. The policy of creating genetic conservation centers for the Syrian subspecies should take into consideration the influence of the oriental lineage O from the northern Syrian border and the large population of genetically divergent indigenous honeybees located in southern Syria.


Preparing for the unknown, responding to the known: communities and public health preparedness.
More than four years after September 11, 2001, bioterrorism preparedness remains a high priority for federal, state, and local governments. With reasonably flexible federal funding, communities have strengthened their ability to respond to public health emergencies, according to assessments by stakeholders and market observers. Collaborative relationships developed for bioterrorism preparedness have proved useful in addressing other threats, such as natural disasters and infectious disease outbreaks. Major ongoing challenges include funding constraints, inadequate surge capacity, public health workforce shortages, competing priorities, and jurisdictional issues.


Injection needles for dental local anesthesia.
The implication of the physical characteristsof single-use disposable dental needles for administration local anesthetics, and the role of these properties in causing breakage, pain on injection, and morbidity in patients are reviewed in this article.


Structure and kinetics of a monomeric glucosamine 6-phosphate deaminase: missing link of the NagB superfamily?
Glucosamine 6-phosphate is converted to fructose 6-phosphate and ammonia by the action of the enzyme glucosamine 6-phosphate deaminase, NagB. This reaction is the final step in the specific GlcNAc utilization pathway and thus decides the metabolic fate of GlcNAc. Sequence analyses suggest that the NagB "superfamily" consists of three main clusters: multimeric and allosterically regulated glucosamine-6-phosphate deaminases (exemplified by Escherichia coli NagB), phosphogluconolactonases, and monomeric hexosamine-6-phosphate deaminases. Here we present the three-dimensional structure and kinetics of the first member of this latter group, the glucosamine-6-phosphate deaminase, NagB, from Bacillus subtilis. The structures were determined in ligand-complexed forms at resolutions around 1.4 Angstroms. BsuNagB is monomeric in solution and as a consequence is active (k(cat) 28 s(-1), K(m(app)) 0.13 mM) without the need for allosteric activators. A decrease in activity at high substrate concentrations may reflect substrate inhibition (with K(i) of approximately 4 mM). The structure completes the NagB superfamily structural landscape and thus allows further interrogation of genomic data in terms of the regulation of NagB and the metabolic fate(s) of glucosamine 6-phosphate.


Catheter ablation vs. antiarrhythmic drugs as 'first-line' initial therapy for atrial fibrillation: a pooled analysis of randomized data.
Catheter ablation (CA) is recommended for patients with atrial fibrillation (AF) after failure of antiarrhythmic drugs (AADs). The role of CA as 'initial therapy' for AF is to be determined. Following PRISMA guideline an up-to-date pooled analysis of randomized data comparing ablation vs. AADs as first-line therapy for symptomatic AF was performed. The primary outcome was recurrence of atrial tachyarrhythmia. The secondary outcomes were improvement in quality-of-life (QoL) and major adverse events. A total of 997 patients from five randomized trials were enrolled (mean age 57.4 years, 68.6% male patients, 98% paroxysmal AF, mean follow-up 1.4 years). The baseline characteristics were similar between the ablation and AADs group. Overall pooled analysis showed that, as compared with AADs, CA as first-line therapy was associated with significantly higher freedom from arrhythmia recurrence (69% vs. 48%, odds ratio: 0.36, 95% confidence interval: 0.27-0.48, P < 0.001). This significance was maintained in subgroup analyses of 1- and 2-year follow-up (P < 0.001). Catheter ablation was associated with significantly greater improvement in QoL regarding AFEQT score and 36-Item Short-Form Health Survey score. The incidence of serious adverse events between ablation and AADs group (5.6% vs. 4.9%, P = 0.62) was similar. Catheter ablation as 'initial therapy' was superior to AADs in maintenance of sinus rhythm and improving QoL for patients with symptomatic paroxysmal AF, without increasing risk of serious adverse events.


Synthesis and structural characterization of the individual diastereoisomers of a cross-stapled alkene-bridged nisin DE-ring mimic.
Herein, we describe the synthesis, structural characterization, and synthetic use as an advanced intermediate of a cross-stapled alkene-bridged hexapeptide to mimic the DE-ring of the lantibiotic nisin. The linear precursor was cyclized by ring-closing metathesis to give the correctly folded bicyclic hexapeptide in a single step, and the four individual diastereoisomers were isolated, structurally assigned and characterized by HPLC, NMR and MS, respectively. The bicyclic hexapeptide was used as a versatile advanced synthon and was modified at its C- and N-terminus, among others, with an azide moiety to access a building block suitable for Cu(I)-catalyzed alkyne-azide cycloaddition-based ligation reactions.


An efficient heterogeneous signcryption for smart grid.
A smart grid, considered the next-generation type of power grid, combines a traditional power grid with information and communication technologies to effectively facilitate power generation and ensure transmission security and reliability in real-time. Only authorized consumers should be able to access the smart grid because the information gathered by smart meters includes users' private information. However, smart grid security is still a challenge. Motivated by this challenge, in this paper, we propose a heterogeneous signcryption (HSC) scheme for secure communication between smart meters and the utility. We demonstrate that this scheme is indistinguishable against adaptive chosen-ciphertext attacks (IND-CCA2), existentially unforgeable against adaptive chosen-message attacks (EUF-CMA) and ciphertext-anonymous against adaptive chosen ciphertext attacks (ANON-CCA2) under the computational Diffie-Hellman (CDH) problem in the random oracle model. Our scheme simultaneously achieves confidentiality, integrity, authentication, non-repudiation and ciphertext anonymity in a single logical step. It supports heterogeneous systems, allowing a meter in an identity-based cryptography (IBC) environment to transmit electrical usage data to a utility in a public key infrastructure (PKI) environment. Compared with other existing related schemes, our scheme has the lowest communication overhead and energy consumption for the smart grid. Based on these features, our scheme is highly suitable for secure power transmissions in a smart grid.


A systems biology approach toward understanding seed composition in soybean.
The molecular, biochemical, and genetic mechanisms that regulate the complex metabolic network of soybean seed development determine the ultimate balance of protein, lipid, and carbohydrate stored in the mature seed. Many of the genes and metabolites that participate in seed metabolism are unknown or poorly defined; even more remains to be understood about the regulation of their metabolic networks. A global omics analysis can provide insights into the regulation of seed metabolism, even without a priori assumptions about the structure of these networks. With the future goal of predictive biology in mind, we have combined metabolomics, transcriptomics, and metabolic flux technologies to reveal the global developmental and metabolic networks that determine the structure and composition of the mature soybean seed. We have coupled this global approach with interactive bioinformatics and statistical analyses to gain insights into the biochemical programs that determine soybean seed composition. For this purpose, we used Plant/Eukaryotic and Microbial Metabolomics Systems Resource (PMR, http://www.metnetdb.org/pmr, a platform that incorporates metabolomics data to develop hypotheses concerning the organization and regulation of metabolic networks, and MetNet systems biology tools http://www.metnetdb.org for plant omics data, a framework to enable interactive visualization of metabolic and regulatory networks. This combination of high-throughput experimental data and bioinformatics analyses has revealed sets of specific genes, genetic perturbations and mechanisms, and metabolic changes that are associated with the developmental variation in soybean seed composition. Researchers can explore these metabolomics and transcriptomics data interactively at PMR.


Tennessee's smokefree policy: it's time for local control.
Tobacco use in Tennessee is higher than the national average and the decline in usage rate has stalled. The smokefree policy enacted to address this health issues contains several exemptions and does not repeal preemption that was introduced in 1994. In March 2013, the Governor unveiled Health and Wellness Initiatives, including reduction in the use of "tobacco products." To achieve this goal, two approaches should be considered and integrated into the initiative to facilitate the decline in tobacco use - policy and population. On the policy approach, the Governor should consider working for the repeal of state preemption of local tobacco control policymaking by the 1994 Prevention of Youth Access to Tobacco Act, and for removing exemption for certain venues from the Non-Smoker Protection Act. On the population approach, the Governor should consider focusing on young adults as an integral part of the target group and tobacco-free campuses as a strategy for addressing tobacco use among such group. All these conform to the Initiatives' strategy of "localized ownership."


A Rare Case of Submucosal Fibroid Presented With Acute Abdomen: A Case Report.
Spontaneous bleeding from uterine leiomyoma is an unusual cause of hemoperitoneum. Fewer than 100 cases have been reported in the literature so far. We report a 30-year-old woman who needed surgical exploration for an acute abdomen. There was massive hemoperitoneum due to bleeding from a subserosal myoma. This case was important because physician should be aware of this rare complication in patient with uterine myoma to avoid any morbidity and mortality.


How blockchain-timestamped protocols could improve the trustworthiness of medical science.
Trust in scientific research is diminished by evidence that data are being manipulated. Outcome switching, data dredging and selective publication are some of the problems that undermine the integrity of published research. Methods for using blockchain to provide proof of pre-specified endpoints in clinical trial protocols were first reported by Carlisle. We wished to empirically test such an approach using a clinical trial protocol where outcome switching has previously been reported. Here we confirm the use of blockchain as a low cost, independently verifiable method to audit and confirm the reliability of scientific studies.


Current trends in the etiology and diagnosis of HPV-related head and neck cancers.
Human papilloma virus (HPV) infection is a major risk factor for a distinct subset of head and neck squamous cell carcinoma (HNSCC). The current review summarizes the epidemiology of HNSCC and the disease burden, the infectious cycle of HPV, the roles of viral oncoproteins, E6 and E7, and the downstream cellular events that lead to malignant transformation. Current techniques for the clinical diagnosis of HPV-associated HNSCC will also be discussed, that is, the detection of HPV DNA, RNA, and the HPV surrogate marker, p16 in tumor tissues, as well as HPV-specific antibodies in serum. Such methods do not allow for the early detection of HPV-associated HNSCC and most cases are at an advanced stage upon diagnosis. Novel noninvasive approaches using oral fluid, a clinically relevant biological fluid, allow for the detection of HPV and cellular alterations in infected cells, which may aid in the early detection and HPV-typing of HNSCC tumors. Noninvasive diagnostic methods will enable early detection and intervention, leading to a significant reduction in mortality and morbidity associated with HNSCC.


Design of a TiO2 nanosheet/nanoparticle gradient film photoanode and its improved performance for dye-sensitized solar cells.
A TiO2 film photoanode with gradient structure in nanosheet/nanoparticle concentration on the fluorine-doped tin oxide glass from substrate to surface was prepared by a screen printing method. The as-prepared dye-sensitized solar cell (DSSC) based on the gradient film electrode exhibited an enhanced photoelectric conversion efficiency of 6.48%, exceeding that of a pure nanoparticle-based DSSC with the same film thickness by a factor of 2.6. The enhanced photovoltaic performance of the gradient film-based DSSC was attributed to the superior light scattering ability of TiO2 nanosheets within the gradient structure, which was beneficial to light harvesting. Furthermore, the TiO2 nanosheets with exposed {001} facets facilitated the electron transport from dye molecules to the conduction band of TiO2 and further to the conductive glass. Meanwhile, the high specific surface area of TiO2 nanosheets helped the adsorption of dye molecules, and the TiO2 nanoparticle underlayer ensured good electronic contact between the TiO2 film and the fluorine-doped tin oxide glass substrate. The electrochemical impedance spectroscopy measurements further confirmed the electron transport differences between DSSCs based on nanosheet/nanoparticle gradient film electrodes and DSSCs based on nanosheet/nanoparticle homogeneous mixtures, pure TiO2 nanoparticles and pure TiO2 nanosheets with the same film thickness.


Biofunctionalized nanoparticles with pH-responsive and cell penetrating blocks for gene delivery.
Bridging the gap between nanoparticulate delivery systems and translational gene therapy is a long sought after requirement in nanomedicine-based applications. However, recent developments regarding nanoparticle functionalization have brought forward the ability to synthesize materials with biofunctional moieties that mimic the evolved features of viral particles. Herein we report the versatile conjugation of both cell penetrating arginine and pH-responsive histidine moieties into the chitosan polymeric backbone, to improve the physicochemical characteristics of the native material. Amino acid coupling was confirmed by 2D TOCSY NMR and Fourier transform infrared spectroscopy. The synthesized chitosan-histidine-arginine (CH-H-R) polymer complexed plasmid DNA biopharmaceuticals, and spontaneously assembled into stable 105 nm nanoparticles with spherical morphology and positive surface charge. The functionalized delivery systems were efficiently internalized into the intracellular compartment, and exhibited remarkably higher transfection efficiency than unmodified chitosan without causing any cytotoxic effect. Additional findings regarding intracellular trafficking events reveal their preferential escape from degradative lysosomal pathways and nuclear localization. Overall, this assembly of nanocarriers with bioinspired moieties provides the foundations for the design of efficient and customizable materials for cancer gene therapy.


Comparison of the efficacy and safety of rosuvastatin versus atorvastatin, simvastatin, and pravastatin across doses (STELLAR* Trial).
The primary objective of this 6-week, parallel-group, open-label, randomized, multicenter trial was to compare rosuvastatin with atorvastatin, pravastatin, and simvastatin across dose ranges for reduction of low-density lipoprotein (LDL) cholesterol. Secondary objectives included comparing rosuvastatin with comparators for other lipid modifications and achievement of National Cholesterol Education Program Adult Treatment Panel III and Joint European Task Force LDL cholesterol goals. After a dietary lead-in period, 2,431 adults with hypercholesterolemia (LDL cholesterol > or =160 and <250 mg/dl; triglycerides <400 mg/dl) were randomized to treatment with rosuvastatin 10, 20, 40, or 80 mg; atorvastatin 10, 20, 40, or 80 mg; simvastatin 10, 20, 40, or 80 mg; or pravastatin 10, 20, or 40 mg. At 6 weeks, across-dose analyses showed that rosuvastatin 10 to 80 mg reduced LDL cholesterol by a mean of 8.2% more than atorvastatin 10 to 80 mg, 26% more than pravastatin 10 to 40 mg, and 12% to 18% more than simvastatin 10 to 80 mg (all p <0.001). Mean percent changes in high-density lipoprotein cholesterol in the rosuvastatin groups were +7.7% to +9.6% compared with +2.1% to +6.8% in all other groups. Across dose ranges, rosuvastatin reduced total cholesterol significantly more (p <0.001) than all comparators and triglycerides significantly more (p <0.001) than simvastatin and pravastatin. Adult Treatment Panel III LDL cholesterol goals were achieved by 82% to 89% of patients treated with rosuvastatin 10 to 40 mg compared with 69% to 85% of patients treated with atorvastatin 10 to 80 mg; the European LDL cholesterol goal of <3.0 mmol/L was achieved by 79% to 92% in rosuvastatin groups compared with 52% to 81% in atorvastatin groups. Drug tolerability was similar across treatments.


Changes in chest CT findings of pulmonary tuberculosis after linezolid treatment.
The purpose of the present study was to evaluate the changes in successive chest CT examinations in patients with pulmonary tuberculosis who achieved culture conversion with linezolid treatment. We reviewed the follow-up CT scans of 14 patients with pulmonary tuberculosis who had sputum-culture conversion after linezolid treatment. This study evaluated cavity, centrilobular nodules, consolidation, bronchial wall thickening, calcified nodule or consolidation, bronchiectasis, irregular lines, and lung destruction. The presence of pleural thickening, pleural effusion and lymphadenopathy was recorded. Follow-up CT scans showed a partial decrease in the extent of centrilobular nodules in all cases. Consolidation was partially cleared in 8 patients and newly developed consolidation was observed in 2 patients. All of the cavities showed a decrease in size and thickness. But the cavities persisted after linezolid treatment in 8 of 9 patients. Bronchial wall thickening was completely or partially cleared in 6 patients and 5 patients, respectively. Newly developed irregular lines, lung destruction and pleural thickening were observed in 1, 1, and 3 patients, respectively. Successive chest CT examinations in patients with linezolid treatment may help in the early assessment of linezolid treatment efficacy because of its rapid availability. Early assessment of linezolid treatment efficacy will help to set up a treatment plan, such as duration of treatment or linezolid dosage. However, they may not be useful for deciding pulmonary tuberculosis activity following linezolid treatment.


Investigation of the effect of PAn and PAn/ZnO photocatalysts on 100% degradation of Congo red under UV visible light irradiation and lightless environment.
PAn (polyaniline) and PAn/ZnO photocatalysts were synthesized using chemical polymerization of aniline. The structure characterization of the synthesized samples was analysed by using X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and UV-Vis spectroscopy measurements, and thermal gravimetric analysis (TGA). The photocatalytic activities of PAn and PAn/ZnO (0.1g/100ml) on the degradation of the Congo red (CR) dye were studied under the UV visible light irradiation and the lightless environment and the efficiency of catalysts have been explained in details. Contribution of UV visible light irradiation on the 100% degradation of CR dye for the PAn and PAn/ZnO photocatalyst is determined. The reaction kinetics and effect of pH (pH 4 and pH 10) were investigated by using first order kinetic model. According to the experimental results, photocatalytic reaction rate of Congo Red increased in acidic environment and under UV visible light irradiation.


Acral Lentiginous Melanomas Harbour Intratumor Heterogeneity in BRAF Exon 15, With Mutations Distinct From V600E/V600K.
The choice of appropriate therapeutic strategies may be influenced by intratumor heterogeneity and makes cancer treatment considerably more challenging. We aimed to evaluate the heterogeneity of BRAF exon 15 mutations in different areas of acral lentiginous melanoma (ALM). The entire exon 15 was sequenced in 4 different areas of paraffin-embedded samples from 26 patients with ALM. A total of 26 of 49 cases of ≥1 mm in depth of ALM identified by clinical, anatomical, and pathological data fulfilled the inclusion and exclusion criteria for this study. Tumors had a mean Breslow depth of 7.2 mm and an average mitotic index of 3 mitosis/mm. Mutations distinct from the common V600E and V600K were detected in 31%, and intratumor heterogeneity was observed in 31% of samples. Interestingly, 63.5% of all mutations had been previously associated with cancer. Most (62.5%) of the missense BRAF exon 15 mutations found in the ALM samples examined here were deemed "detrimental" for protein function according to at least 2 functional prediction programs, and 3 mutations (37.5%) were predicted to be "neutral," with no effect on protein function. BRAF exon 15 mutations were detected frequently in ALM and displayed heterogeneity, a finding to be further investigated.


C242T CYBA polymorphism of the NADPH oxidase is associated with reduced respiratory burst in human neutrophils.
Oxidative stress contributes to the pathogenesis of atherosclerosis. p22phox-based NAD(P)H oxidases exist in the vessel wall, acting as important superoxide-generating systems in the vasculature. Some studies have identified reduced atherosclerosis in the presence of the C242T CYBA polymorphism, whereas others have not. Because vascular p22phox is identical to neutrophil p22phox, we studied the association between the C242T, A640G, and -930A/G CYBA polymorphisms and the quantity of superoxide produced from neutrophils isolated from healthy adults to determine if these polymorphisms had any functional impact on NADPH oxidase function. Neutrophils were isolated from 90 subjects by Percoll density gradient centrifugation. Genotypes were determined by polymerase chain reaction (PCR) and restriction mapping, as well as real-time PCR. The oxidative burst was stimulated with phorbol 12-myristate 13-acetate. Superoxide was quantified using the superoxide dismutase inhibitable oxidation of the spin probe hydroxylamine 1-hydroxy-3-carboxy-pyrrolidine, detected by electron paramagnetic resonance. Superoxide production was significantly affected by the C242T polymorphism, being 8.7+/-0.7, 7.9+/-0.6, and 5.9+/-1.2 micromol/L per minute per 10(6) neutrophils for the C242T CC, CT, and TT genotypes, respectively (P<0.05). In contrast, the A640G and the -930A/G polymorphisms did not alter the neutrophil respiratory burst. Phagocytic respiratory burst activity in homozygous individuals with the T allele of the C242T CYBA polymorphism is significantly lower than of wild-type carriers and heterozygous individuals. Because p22phox exists in both the neutrophil and vessel wall, vascular oxidative stress is likely diminished in individuals with this polymorphism.


Ecological determinants and temporal stability of the within-river population structure in Atlantic salmon (Salmo salar L.).
A gene diversity analysis was performed using microsatellite loci in order to (i) describe the extent and pattern of population structure in Atlantic salmon (Salmo salar L.) within a river system; (ii) establish the importance of quantifying the signal:noise ratio in accurately estimating population structure; and (iii) assess the potential usefulness of two evolutionary models in explaining within-river population structure from the ecological and habitat characteristics of Atlantic salmon. We found weak, yet highly significant microscale spatial patterning after accounting for variance among temporal replicates within sites. Lower genetic distances were observed among temporal samples at four sampling sites whereas no evidence for temporal stability was observed at the other three locations. The component of genetic variance attributable to either temporal instability and/or random sampling errors was almost three times more important than the pure spatial component. This indicates that not considering signal:noise ratio may lead to an important overestimation of genetic substructuring in situations of weak genetic differentiation. This study also illustrates the usefulness of the member-vagrant hypothesis to generate a priori predictions regarding the number of subpopulations that should compose a species, given its life-history characteristics and habitat structure. On the other hand, a metapopulation model appears better suited to explain the extent of genetic divergence among subpopulations, as well as its temporal persistence, given the reality of habitat patchiness and environment instability. We thus conclude that the combined use of both models may offer a promising avenue for studies aiming to understand the dynamics of genetic structure of species found in unstable environments.


[Values, ethical positions and attitudes of candidates for a lawyer career].
This work intends to bring out light regarding values, ethical positions and attitudes of future Law students. We consider that it may be important to know these aspects in relation to the vocational decision of students in order to know whether values are really a determinant factor in career decision or not. The present study was produced using the following instruments: Study of Values (Allport-Vernon and Lindzey); Survey of Interpersonal Values (L. Gordon); Ethical Position Questionnaire (J. E. Moreno). Several groups of students were used as samples: neophytes to Law School in the University of Buenos Aires, and neophytes to the careers of Law, Medicine, Psychology and Publicity in a Catholic University. We found that in the latter University the predominant value is the religious, and secondly the social value. Instead in the University of Buenos Aires the most important value is the social and secondly the political value. The EPQ shows that neophytes to Law School may be considered absolutists in Forsyth's scale. These students also showed in Gordon's survey high Benevolence and Conformity in comparison with the other samples of students. Regarding attitudes towards the University's objectives, we found that Law students consider important the ethical education and the enhancement of the national culture more than universal culture. This work tends to confirm that Law neophytes as a group have clear differences from other groups of neophytes regarding attitudes to interpersonal bonds, and also different attitudes toward the university's objectives.


Behaviour of biofilm systems under varying hydrodynamic conditions.
Heterotrophic biofilms were cultivated in long-term experiments in biofilm tube reactors. During the biofilm cultivation the substrate loading of glucose was kept constant while the hydrodynamic conditions were changed stepwise. To describe the behaviour of the biofilm structure under these varying flow conditions the mass transfer and transport at the bulk/biofilm interface and inside the biofilm was investigated with oxygen microelectrodes. Furthermore, the biofilm density was used to describe the biofilm compactness before and after the change of the hydrodynamic condition. The obtained results show that the biofilm density and also the substrate flux decreased with decreasing flow velocity in the bulk phase. Additionally the slope of the oxygen concentration profiles decreased and the thickness of the concentration boundary layer increased. On the other hand, increasing the flow velocity in the bulk phase led both to a higher biofilm density and a higher maximum substrate flux. The biofilm surface became more homogenous and the thickness of the concentration boundary layer decreased. The time for adaptation of the biofilm structure after changing the hydrodynamic conditions ranged between 1 and 3 weeks.


ITV versus mid-ventilation for treatment planning in lung SBRT: a comparison of target coverage and PTV adequacy by using in-treatment 4D cone beam CT.
The internal target volume (ITV) approach and the mid-ventilation (MidV) concept are the two main respiratory motion-management strategies under free breathing. The purpose of this work was to compare the actual in-treatment target coverage during volumetric modulated arctherapy (VMAT) delivered through both ITV-based and MidV-based planning target volume (PTV) and to provide knowledge in choosing the optimal PTV for stereotactic body radiotherapy (SBRT) for lung lesions. Thirty-two lung cancer patients treated by a VMAT technique were included in the study. For each fraction, the mean time-weighted position of the target was localized by using a 4-dimensional cone-beam CT (4D-CBCT)-based image guidance procedure. The respiratory-correlated location of the gross tumor volume (GTV) during treatment delivery was determined for each fraction by using in-treatment 4D-CBCT images acquired concurrently with VMAT delivery (4D-CBCTin-treat). The GTV was delineated from each of the ten respiratory phase-sorted 4D-CBCTin-treat datasets for each fraction. We defined target coverage as the average percentage of the GTV included within the PTV during the patient's breathing cycle averaged over the treatment course. Target coverage and PTVs were reported for a MidV-based PTV (PTVMidV) using dose-probabilistic margins and three ITV-based PTVs using isotropic margins of 5 mm (PTVITV + 5mm), 4 mm (PTVITV + 4mm) and 3 mm (PTVITV + 3mm). The in-treatment baseline displacements and target motion amplitudes were reported to evaluate the impact of both parameters on target coverage. Overall, 100 4D-CBCTin-treat images were analyzed. The mean target coverage was 98.6, 99.6, 98.9 and 97.2% for PTVMidV, PTVITV + 5mm, PTVITV + 4mm and PTVITV + 3mm, respectively. All the PTV margins led to a target coverage per treatment higher than 95% in at least 90% of the evaluated cases. Compared to PTVITV + 5mm, PTVMidV, PTVITV + 4mm and PTVITV + 3mm had mean PTV reductions of 16, 19 and 33%, respectively. When implementing VMAT with 4D-CBCT-based image guidance, an ITV-based approach with a tighter margin than the commonly used 5 mm margin remains an alternative to the MidV-based approach for reducing healthy tissue exposure in lung SBRT. Compared to PTVMidV, PTVITV + 3mm significantly reduced the PTV while still maintaining an adequate in-treatment target coverage.


Intake of Nutrients, Fiber, and Sugar in Patients with Nonalcoholic Fatty Liver Disease in Comparison to Healthy Individuals.
Nonalcoholic fatty liver disease (NAFLD) is the most common cause of chronic liver disease in the world. Although some studies have been conducted about dietary intakes of these patients, but the results are inconsistent. The aim of this study was to survey all macronutrients and micronutrients included in dietary intake of these patients for better understanding the factors influencing this disease. The present study is a case-control conducted in Isfahan city, Iran. The cases were recently diagnosed patients with NAFLD who identified by ultrasonography. The case (159) and control (158) individuals were matched in age and gender. Data of general characteristics and physical activity of individuals were collected through questionnaire. Dietary intake was also collected using 24 h dietary recall questionnaire. Waistline and body mass index for the case group were more than the control group (P < 0.05). Physical activity level in healthy individuals was more than patients with NAFLD. Dietary intake of saturated fatty acids and sugar in patients with NAFLD was more than healthy individuals (P < 0.05). Intake of total dietary fiber, folic acid, Vitamin D, zinc, and potassium in healthy individuals was more than patients with NAFLD (P < 0.05). In total, it seems the type of dietary intake source is associated with NAFLD. Increasing saturated fatty acids and sugar and decreasing fiber, folic acid, Vitamin D, zinc, and potassium intake might play a role in the progression of this disease.


Continuous glucose monitoring in older people with diabetes receiving home care-a feasibility study.
Hypoglycemic incidents in older people can cause severe health problems, enhance general age-related disabilities, and cause frailty. Little is known about incidences of hypoglycemia in older home-dwelling people with diabetes. Thus, the aim of this study was to examine the feasibility of capturing hypoglycemia and issues associated with increased risk of hypoglycemia by use of continuous glucose monitoring (CGM) and standardized questionnaires among older home-dwelling individuals with diabetes type 2 receiving home care. CGM with the Ipro2-blinded monitoring system were performed for 5 days in six home-dwelling individuals ≥ 75 years diagnosed with diabetes and receiving home care. Demographic (age, gender, living arrangements) and clinical data (diabetes diagnoses and duration, diabetes medication, documented treatment goal, available glycosylated hemoglobin (HbA1c)) were collected from electronic patient records in home care services. Feasibility (ease of use, quality of data, time spent) of standardized questionnaires to identify the risk of hypoglycemia (the McKellar Risk Assessment Tool), risk of malnutrition (the Mini Nutritional Assessment (MNA)), functional status (the Individual-based Statistics for Nursing and Care Services (IPLOS)), and cognitive status (the Mini Mental Status Exam (MMSE)) was also assessed. Questionnaire data was collected by a study nurse in the individuals' home. The practical use of CGM was satisfactory, with no major remarks about discomfort or technical errors, except for one participant with skin reaction (redness). Collecting data with the McKellar Risk Assessment Tool, MNA and IPLOS worked well according to quality of data, time spent, and ease of use. The MMSE survey required extensive training of personnel to be conducted. The feasibility study informs an upcoming study on the incidence and risk factors of hypoglycemia in home-dwelling older individuals. We will ascertain that personnel who will use the MMSE questionnaire to collect cognitive status and skills are familiar with the tool and adequately educated and trained before study start. The use of blinded CGM in this population was well tolerated and can be used "as is" for future studies.